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FRS2 family docking proteins with overlapping roles in activation of MAP kinase have distinct spatial-temporal patterns of expression of their transcripts.

FRS2alpha and FRS2beta, two members of the FRS2 family of docking proteins, become tyrosine phosphorylated in response to fibroblast growth factor (FGF) or nerve growth factor (NGF) stimulation. Tyrosine phosphorylated FRS2alpha serves as a platform for the recruitment of multiple signaling proteins for activation of the Ras-mitogen-activated protein (MAP) kinase signaling cascade. We report that Frs2alpha and Frs2beta have distinct spatio-temporal expression patterns in mouse embryos. We further show that FRS2beta can compensate for the loss of FRS2alpha for activation of MAP kinase when expressed in fibroblasts from Frs2alpha(-/-) mouse embryos. We propose that the FRS2 family proteins have distinct roles in vivo through activation of common signaling proteins including MAP kinase.

Adaptor Proteins, Signal Transducing↗

Spatial heterogeneity of aggregate stability and soil carbon in semi-arid rangeland.

To measure and manage for C sequestration in heterogeneous rangeland systems, we need to more fully understand spatial patterns of soil resources. Spatial distributions of aggregate stability and soil carbon were investigated in a semiarid rangeland in New Mexico, USA. Soil was analyzed from plant interspaces, black grama (Bouteloua eriopoda (Torr.) Torr.), and mesquite (Prosopis glandulosa Torr.) in a landscape-replicated study. Aggregate stability at the 250 microm scale, carbonate C, organic C and N, C:N ratio, and glomalin, were all highest under mesquite. Soil C:N ratio was the best predictor of aggregate stability. Estimates of metric tons of C per hectare in the top 10 cm were highly variable at patch and landscape scales, varying from 4.2 to 10.5 under mesquite and from 3.0 to 7.0 in interspaces. High variability of aggregate stability and soil C has important implications for C sequestration. We argue that this multi-scale soil heterogeneity must be considered when measuring and managing for C sequestration.

Carbon↗

Memorizing and recalling spatial-temporal patterns in an oscillator model of the hippocampus.

We describe the model of the hippocampus consisting of interactive oscillators with input from the entorhinal cortex (modulating the main information flow by a theta rhythm) and the septum (a theta rhythm generator). When interconnections between oscillators are allowed to strengthen in an adaptive way, the network can be trained using a series of lessons. This results in a connection matrix that memorizes the temporal sequence of inputs. Presenting one of the lessons to the trained network results in reproduction of the remainder of the sequence. In this paper, we create such a connection matrix, derive from it an appropriate Markov chain and simulate the chain to illustrate its dynamics.

Hippocampus↗

Spatial cortical patterns of metabolic activity in monkeys performing a visually guided reaching task with one forelimb.

The 2-[14C]deoxyglucose method was used to map the metabolic activity in the neocortex of monkeys (Macaca nemestrina) performing a visually guided reaching task with one forelimb. Monkeys received liquid reward for correct, single directional reaching movements, which were required at a rate of about 10 per minute. We estimated the weighted average of local glucose consumption within several neocortical areas, and we reconstructed quantitative, high-resolution, two-dimensional maps of the detailed spatiointensive patterns of activity. Our findings demonstrate the involvement of the striate and prestriate cortices, the inferior intraparietal and superior temporal visual association areas, the frontal eye field and the caudal periprincipal cortex, the primary somatosensory and the related superior intraparietal area, the primary and association auditory cortices, the superior temporal multimodal region, and the premotor, primary, supplementary, and cingulate motor areas. The visual cortex in the superior temporal and the intraparietal sulci, which is concerned with "where", was activated during visually guided reaching. In contrast, the inferior temporal visual association cortex, which is concerned with "what", was not involved in our study. We suggest that the activated direction-selective layer four of V1 and the thick stripes of V2 convey visuomotor information to the activated cortex in the posterior bank and the floor of the superior temporal sulcus, which may encode the constantly updated position of the moving forelimb. In parallel, the activated cortex in the ventral part and the lateral bank of the intraparietal sulcus may encode visuospatial information related to the localization of the visual target in the extrapersonal space. Furthermore, the dorsal part of the medial bank of the intraparietal sulcus may be involved in proprioceptive guidance of movement, based on the parallel metabolic effects shown only contralateral to the moving forelimb within this region and the forelimb representations in the primary somatosensory and motor cortices. Finally, the bilaterally activated network including the inferior postarcuate skeletomotor and prearcuate oculomotor cortical fields and the caudal periprincipal region 46 may participate in sensory and oculomotor to motor transformations, in parallel with the medial and lateral intraparietal cortices with which this network is reciprocally interconnected.

Animals↗

Spatial expression patterns of epidermal growth factor receptor gene transcripts in the postnatal mammalian cochlea.

Recent in vitro studies demonstrated that members of the epidermal growth factor (EGF) family are involved in hair cell replacement in the postnatal mammalian organ of Corti (OC) after ototoxic damage. This suggests a role for the EGF receptor (EGFR) in this process. We examined the expression of EGFR mRNA within the normal postnatal day 3 (P3) and adult rat cochlear epithelium by RT-PCR and examined its cellular localization with non-radioactive in situ hybridization in P3 and adult cochleae. RT-PCR demonstrated that EGFR mRNA is expressed in P3 and adult cochlear epithelium. In situ hybridization localized high levels of EGFR transcripts in the OC, spiral ganglion, Kölliker's organ and detectable levels in the supporting cells and the stria vascularis of P3 cochlea. In the adult cochlea, EGFR transcripts were detected only in the spiral ganglion. Our results support that the EGFR is implicated in the differentiation of several cochlear cell types and in the response of OC to ototoxic damage of the P3 rat. In the adult, it may participate in the maintenance of the mature neurons and its absence in the OC may contribute to the lack of regenerative responses in the adult cochlea.

Animals↗

Temporal and spatial expression patterns of FoxD2 during the early development of Xenopus laevis.

We have investigated the sequence and expression pattern of the Xenopus laevis FoxD2 gene, a member of the fork head/winged helix multigene family. The derived protein sequence is most closely related to FoxD2 factors known from other species. Maternal FoxD2 transcripts are degraded during early cleavage stages. Zygotic transcription is activated after the midblastula transition followed by a pronounced increase during neurulation. Whole mount in situ hybridisations reveal that FoxD2 is predominantly expressed in the paraxial mesoderm, but not within the myotome. In addition, FoxD2 transcripts are found within the migrating ventral abdominal muscle precursors, in cranial neural crest cells surrounding the eye and populating the second and third visceral arches as well as within restricted areas of the diencephalon. In hatched tadpoles, FoxD2 expression is also observed within the terminal part of the gut.

Amino Acid Sequence↗

A graded response to BMP-4 spatially coordinates patterning of the mesoderm and ectoderm in the zebrafish.

The effects of signal perturbation on expression domains of molecular markers for the mesoderm and ectoderm have been analysed across the dorso-ventral axis in zebrafish embryos. Injection of RNA encoding bone morphogenetic protein-4 (BMP-4) ventralised the embryo, expanding the intermediate mesoderm and non-neural ectoderm at the expense of the dorso-anterior mesoderm and neural plate. A dose-dependent response was observed both morphologically and in expression of gta3, MyoD and pax2. Conversely, increases in dorso-anterior mesoderm and neurectoderm were generated by injection of RNA encoding either a dominant-negative BMP receptor (delta BMPR) or noggin, as demonstrated by goosecoid and pax2 expression. Ventral BMP-4 expression was also inhibited. Thus, patterning of both the mesoderm and the ectoderm during gastrulation appears to depend, directly or indirectly, on the level of BMP activity. Consistent with their locations prior to formation of the neural tube, elevated BMP-4 increased the number of dorsal spinal cord neurons whilst sonic hedgehog and islet1 expression in the ventral spinal cord were reduced. However, the ectopic neurons were not positioned more ventrally, implicating a prepattern in the dorsal neural tube that is independent of the ventral central nervous system.

Animals↗

The temporal and spatial transcription pattern in root nodules of Vicia faba nodulin genes encoding glycine-rich proteins.

Four different transcript sequences encoding gene products with an unusually high glycine content were identified in Vicia faba root nodules. Northern blot analysis revealed a strong nodule specific expression of the corresponding genes. Time course experiments showed that two of these genes were transcribed before the onset of leghemoglobin expression and hence were designated VfENOD-GRP2 and VfENOD-GRP5, whereas the first detection of VfNOD-GRP1 and VfNOD-GRP4 transcripts coincided with the appearance of leghemoglobin transcripts in V. faba root nodules. A characteristic feature of all encoded nodulins was a hydrophobic N-terminus, which in the case of the nodulins ENOD-GRP2 and ENOD-GRP5 has the characteristics of a signal peptide. Such a structure is comparable to other plant glycine-rich proteins decribed as components of the plant cell wall. Based on tissue print hybridizations, we found that VfNOD-GRP1, VfENOD-GRP2 and VfNOD-GRP4 were expressed in the interzone II-III and in the whole nitrogen-fixing zone III. In contrast to VfENOD-GRP2 and VfNOD-GRP4, the signal intensity of hybridizing VfNOD-GRP1 transcripts was slightly reduced in the more proximal part of broad bean root nodules. Apart from the interzone II-III and the nitrogen fixing zone III, VfENOD-GRP5 RNA was also detected in large areas of the prefixing zone II.

Amino Acid Sequence↗

Comparison of ventral subicular and hippocampal neuron spatial firing patterns in complex and simplified environments.

Activity from ventral subicular and hippocampal CA1 neurons was recorded in rats exploring a 4-arm radial maze in which the local and distal cues could be manipulated. Cells from both regions exhibited place fields, although ventral subicular neurons had larger fields than hippocampal cells. Rotation of the local and distal cues in opposite directions produced movement of the place fields in either direction or a complete change in firing pattern. Simplifying the environment also produced changes in place field location. Despite similarities between regions, subiculum fields decreased in size whereas hippocampal fields increased in the simple environment. These findings suggest that subicular cells may receive converging input from several hippocampal neurons and code more complex configurations of the cues.

Analysis of Variance↗

Spatial-temporal patterns of gene expression in mouse skeletal muscle after injection of lacZ plasmid DNA.

Gene therapy for muscular diseases requires the efficient transfection of a large proportion of myofiber cells within a given muscle. In the present experiments, patterns of beta-galactosidase expression were examined in mouse rectus femoris muscles at various time-points after a single injection of lacZ encoded plasmid DNA. beta-Galactosidase expression was detected 3 h after injection and rose to peak levels at 3-14 days, and then stabilized at lower levels. beta-Galactosidase staining was detected in an average of about 6% (up to 15%) of the total 4000 myofiber cells, and in about 70% of those myofibers located in the discrete area containing the greatest proportion of transfected cells. Soon after injection of DNA encoding cytoplasmic or nuclear-targeted beta-galactosidase, expression was noted predominantly in the myotendinous junction areas, after which beta-galactosidase activity progressed toward the central parts of the myofibers. This preferential transgene expression at the myotendinous junction may result from some unique, local property of the myofiber cells and/or from a restricted diffusion or binding of the injected plasmid DNA at tendinous surfaces. A better understanding of the reasons for this pattern of reporter gene expression in muscle may suggest procedures for increasing the number of myofiber cells transfected by direct DNA injections.

Animals↗

A cotyledon regulatory region is responsible for the different spatial expression patterns of Arabidopsis 2S albumin genes.

The 2S albumin genes of Arabidopsis thaliana are a model system to study gene expression during late embryogenesis. The at2S1 gene has previously been shown to be expressed essentially in the embryo axis, unlike at2S2, which is expressed throughout the embryo. Hybrid promoter constructs between at2S1 and at2S2 were introduced into Arabidopsis and used to identify a cotyledon regulatory region necessary for 2S albumin expression in palisade parenchyma and specific epidermal cells. Other promoter sequences flanking this tissue-specific promoter element were shown to control mRNA expression levels independently of the mRNA distribution throughout the embryos. Certain hybrid promoters resulted in the alteration of the time course of expression in cotyledons. Differential expression of 2S albumin genes is discussed in terms of layered cellular organization and mitotic activity throughout the embryo.

Albumins↗

Spatial-temporal patterning of metabotropic glutamate receptor-mediated inositol 1,4,5-triphosphate, calcium, and protein kinase C oscillations: protein kinase C-dependent receptor phosphorylation is not required.

The metabotropic glutamate receptors (mGluR), mGluR1a and mGluR5a, are G protein-coupled receptors that couple via G(q) to the hydrolysis of phosphoinositides, the release of Ca(2+) from intracellular stores, and the activation of protein kinase C (PKC). We show here that mGluR1/5 activation results in oscillatory G protein coupling to phospholipase C thereby stimulating oscillations in both inositol 1,4,5-triphosphate formation and intracellular Ca(2+) concentrations. The mGluR1/5-stimulated Ca(2+) oscillations are translated into the synchronized repetitive redistribution of PKCbetaII between the cytosol and plasma membrane. The frequency at which mGluR1a and mGluR5a subtypes stimulate inositol 1,4,5-triphosphate, Ca(2+), and PKCbetaII oscillations is regulated by the charge of a single amino acid residue localized within their G protein-coupling domains. However, oscillatory mGluR signaling does not involve the repetitive feedback phosphorylation and desensitization of mGluR activity, since mutation of the putative PKC consensus sites within the first and second intracellular loops as well as the carboxyl-terminal tail does not prevent mGluR1a-stimulated PKCbetaII oscillations. Furthermore, oscillations in Ca(2+) continued in the presence of PKC inhibitors, which blocked PKCbetaII redistribution from the plasma membrane back into the cytosol. We conclude that oscillatory mGluR signaling represents an intrinsic receptor/G protein coupling property that does not involve PKC feedback phosphorylation.

Calcium↗

Spatial expression patterns and biochemical properties distinguish a second myo-inositol monophosphatase IMPA2 from IMPA1.

Lithium is used in the clinical treatment of bipolar disorder, a disease where patients suffer mood swings between mania and depression. Although the mode of action of lithium remains elusive, a putative primary target is thought to be inositol monophosphatase (IMPase) activity. Two IMPase genes have been identified in mammals, the well characterized myo-inositol monophosphatase 1 (IMPA1) and myo-inositol monophosphatase 2 (IMPA2). Several lines of genetic evidence have implicated IMPA2 in the pathogenesis of not only bipolar disorder but also schizophrenia and febrile seizures. However, little is known about the protein, although it is predicted to have lithium-inhibitable IMPase activity based on its homology to IMPA1. Here we present the first biochemical study comparing the enzyme activity of IMPA2 to that of IMPA1. We demonstrate that in vivo, IMPA2 forms homodimers but no heterodimers with IMPA1. Recombinant IMPA2 exhibits IMPase activity, although maximal activity requires higher concentrations of magnesium and a higher pH. IMPA2 shows significantly lower activity toward myo-inositol monophosphate than IMPA1. We therefore screened for additional substrates that could be more efficiently dephosphorylated by IMPA2, but failed to find any. Importantly, when using myo-inositol monophosphate as a substrate, the IMPase activity of IMPA2 was inhibited at high lithium and restricted magnesium concentrations. This kinetics distinguishes it from IMPA1. We also observed a characteristic pattern of differential expression between IMPA1 and IMPA2 in a selection of tissues including the brain, small intestine, and kidney. These data suggest that IMPA2 has a separate function in vivo from that of IMPA1.

Amino Acid Sequence↗

The genetic diversity of two brazilian vellozia (velloziaceae) with different patterns of spatial distribution and pollination biology.

BACKGROUND AND AIMS: The genetic structure and variability of two species of Vellozia (Velloziaceae) with restricted distribution in high-altitude quartzitic fields in south-eastern Brazil were studied. Vellozia epidendroides is short, grows on pebbly or sandy soil, and is pollinated by bees. Vellozia leptopetala is arborescent, grows on rock outcrops, and is pollinated by bees and hummingbirds. Both are self-incompatible and have a short, massive flowering strategy. The study aimed to associate differences in their genetic diversity and structure with their microhabitat distribution and pollination ecology. METHODS: Leaves from 106 and 139 plants of V. epidendroides and V. leptopetala, respectively, were collected from five patches of each species and prepared for electrophoretic analyses. KEY RESULTS: Five enzyme systems could be reliably scored for both species. Vellozia epidendroides showed 100 % of the loci polymorphic for almost all patches. The average number of alleles per locus ranged between 2.2 and 2.4 among patches. The Wright's fixation index (F) for this species was 0.226. A significant (p) value indicates that there is a reasonable genetic divergence among patches. Vellozia leptopetala presented 47.5 % of polymorphic loci. All levels of P, A, A(p) and of heterozygosities were lower than those of V. epidendroides. Vellozia leptopetala showed high inbreeding within patches. CONCLUSIONS: The relatively high values of genetic diversity indices found for V. epidendroides may be associated with its large and widespread populations. On the other hand, the low values of genetic diversity found for V. leptopetala may be related to physical isolation on outcrops and intensive foraging by territorial hummingbirds, which may hinder gene flow among patches, aggravated by the very restricted seed dispersal characteristic of the genus, that facilitates sibling mating. It is important to stress the need to preserve the specific habitats of these species of Vellozia, in particular those of V. leptopetala that has lower genetic diversity and is restricted to rock outcrop environments.

Brazil↗

Aberrant splicing and altered spatial expression patterns in fruitless mutants of Drosophila melanogaster.

The fruitless (fru) gene functions in Drosophila males to establish the potential for male sexual behaviors. fru encodes a complex set of sex-specific and sex-nonspecific mRNAs through the use of multiple promoters and alternative pre-mRNA processing. The male-specific transcripts produced from the distal (P1) fru promoter are believed to be responsible for its role in specifying sexual behavior and are only expressed in a small fraction of central nervous system (CNS) cells. To understand the molecular etiology of fruitless mutant phenotypes, we compared wild-type and mutant transcription patterns. These experiments revealed that the fru(2), fru(3), fru(4), and fru(sat) mutations, which are due to P-element inserts, alter the pattern of sex-specific and sex-nonspecific fru RNAs. These changes arise in part from the P-element insertions containing splice acceptor sites that create alternative processing pathways. In situ hybridization revealed no alterations in the locations of cells expressing the P1-fru-promoter-derived transcripts in fru(2), fru(3), fru(4), and fru(sat) pharate adults. For the fru(1) mutant (which is due to an inversion breakpoint near the P1 promoter), Northern analyses revealed no significant changes in fru transcript patterns. However, in situ hybridization revealed anomalies in the level and distribution of P1-derived transcripts: in fru(1) males, fewer P1-expressing neurons are found in regions of the dorsal lateral protocerebrum and abdominal ganglion compared to wild-type males. In other regions of the CNS, expression of these transcripts appears normal in fru(1) males. The loss of fruitless expression in these regions likely accounts for the striking courtship abnormalities exhibited by fru(1) males. Thus, we suggest that the mutant phenotypes in fru(2), fru(3), fru(4), and fru(sat) animals are due to a failure to appropriately splice P1 transcripts, whereas the mutant phenotype of fru(1) animals is due to the reduction or absence of P1 transcripts within specific regions of the CNS.

Amino Acid Sequence↗

Faces evoke spatially differentiated patterns of BOLD activation and deactivation.

Using fMRI techniques sensitive to blood oxygen-level dependent (BOLD) contrast, we measured brain activity in participants (n=8) as they viewed images of faces presented periodically within a continuously changing montage of common objects. Consistent with prior studies, we identified regions of ventral extrastriate cortex, primarily in the fusiform and inferior temporal gyri and nearby cortex, that were activated by faces as measured by an increase in BOLD signal. In addition, we made the novel observation that faces deactivated other areas of ventral extrastriate cortex, primarily in the lingual and parahippocampal gyri and medial to activations. These deactivated regions, identified by a decrease in BOLD signal, may reflect populations of neurons that decrease their activity when faces appear, possibly as a consequence of category-specific inhibition.

Adult↗